Recent advancements in single-cell metabolic analysis for pharmacological research

Journal of Pharmaceutical Analysis, Journal Year: 2023, Volume and Issue: 13(10), P. 1102 - 1116

Published: Aug. 23, 2023

Cellular heterogeneity is crucial for understanding tissue biology and disease pathophysiology. Pharmacological research being advanced by single-cell metabolic analysis, which offers a technique to identify variations in RNA, proteins, metabolites, drug molecules cells. In this review, the recent advancement of analysis techniques their applications metabolism response are summarized. High-precision controlled isolation manipulation provided microfluidics-based methods, such as droplet microfluidics, microchamber, open microfluidic probe, digital microfluidics. They used tandem with variety detection techniques, including optical imaging, Raman spectroscopy, electrochemical detection, RNA sequencing, mass spectrometry, evaluate changes administration. The advantages disadvantages different discussed along challenges future directions analysis. These employed pharmaceutical studying resistance pathway, therapeutic targets discovery, vitro model evaluation.

Language: Английский

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Single-cell and bulk transcriptional profiling of mouse ovaries reveals novel genes and pathways associated with DNA damage response in oocytes.

Developmental Biology, Journal Year: 2024, Volume and Issue: 517, P. 55 - 72

Published: Sept. 19, 2024

Language: Английский

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Single-cell RNA sequencing analysis of mouse follicular somatic cells

Biology of Reproduction, Journal Year: 2021, Volume and Issue: 105(5), P. 1234 - 1245

Published: Aug. 31, 2021

Within the development of ovarian follicle, in addition to cell proliferation and differentiation, sophisticated cell-cell cross talks are established among follicular somatic cells such as granulosa (GCs) theca cells. To systematically reveal differentiation signal transductions cells, we collected mouse from secondary ovulatory stage, analyzed single transcriptomes. Having data filtered screened, found 6883 high variable genes 4888 Then were clustered into 26 clusters, including 18 GC 4 endocrine (TEC) other which include immune Acta2 positive externa From our data, there was metabolic reprogramming happened during differentiation. We also both Cyp19a1 Cyp11a1 could be expressed TECs. expression patterns associated with interactions steroid hormone receptor genes, insulin signaling cytokine/transformation growth factor beta all clusters. Lastly, highly 300 gene used search new involved follicle development. These transcriptomes provide us potential clues how mammals regulating help find targets improve oocyte quality for women low fertility.

Language: Английский

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Single-nucleus RNA Sequencing reveals the mechanism of cigarette smoke exposure on diminished ovarian reserve in mice

Ecotoxicology and Environmental Safety, Journal Year: 2022, Volume and Issue: 245, P. 114093 - 114093

Published: Sept. 16, 2022

The systematic toxicological mechanism of cigarette smoke (CS) on ovarian reserve has not been extensively investigated. Female 8-week-old C57BL/6 mice at peak fertility were exposed to CS or indoor air only for 30 days (100 per group) and the effects assessed using Single-Nucleus RNA Sequencing (snRNA-seq). In addition, further biochemical experiments, including immunohistochemical staining, ELISA, immunofluorescence transmission electron microscopy, cell counting kit-8 assay, flow cytometry analysis, senescence-associated β-galactosidase western blotting, accomplished confirm snRNA-seq results. We identified nine main types in adult ovaries cell-type-specific differentially expressed genes (DEGs) induced by exposure. Western blot results verified that down-regulation antioxidant (Gpx1 Wnt10b) steroid biosynthesis gene (Fdx1) occurred both tissue human granulosa cell-like tumor line (KGN cells) after Five percent extract (CSE) effectively stimulated production reactive oxygen species (ROS), DNA damage, cellular senescence markedly inhibited KGN proliferation inducing G1-phase cycle arrest. Moreover, Gja1, Lama1 Ferroptosis indicator (Gpx4) cells plays a significant role ultrastructural changes ovary These observations suggest exposure impaired follicle might be caused REDOX imbalance cells. current study systematically determined damage mouse provides theoretical basis early clinical prediction, diagnosis intervention exposure-associated primary insufficiency (POI), is great significance improving female reproductive health.

Language: Английский

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Single-Cell Transcriptomics Analysis Reveals a Cell Atlas and Cell Communication in Yak Ovary

International Journal of Molecular Sciences, Journal Year: 2023, Volume and Issue: 24(3), P. 1839 - 1839

Published: Jan. 17, 2023

Yaks (Bos grunniens) are the only bovine species that adapt well to harsh high-altitude environment in Qinghai-Tibetan plateau. However, reproductive adaptation climate of high elevation remains be elucidated. Cell composition and molecular characteristics foundation normal ovary function which determines performance. So, delineating ovarian at a cellular level is conducive elucidating mechanism underlying adaption yaks. Here, single-cell RNA-sequencing (scRNA-seq) was employed depict an atlas containing different cell types with specific signatures yak ovary. The were identified on basis their specifically expressed genes biological functions. As result, established successfully theca cells, stromal endothelial smooth muscle natural killer macrophages, proliferating cells. A cell-to-cell communication network between distinct constructed. cells clustered into five subtypes based Further, CYP11A1 confirmed as marker gene for by immunofluorescence staining. Our work reveals contributes providing insights

Language: Английский

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AMH protects the ovary from doxorubicin by regulating cell fate and the response to DNA damage

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(5)

Published: Jan. 28, 2025

Anti-Müllerian hormone (AMH) protects the ovarian reserve from chemotherapy, and this effect is most pronounced with Doxorubicin (DOX). However, DOX toxicity AMH rescue mechanisms in ovary have remained unclear. Herein, we characterize consequences of these treatments cell types using scRNAseq. DOX-induced DNA damage activates Tp53 class mediators across types. In mesenchyme, cotreatment halts theca progenitor differentiation reduces apoptotic gene expression. preantral granulosa cells, upregulates cycle inhibitor Cdkn1a dysregulates Wnt signaling, which are ameliorated by cotreatment. Finally, induces Id3 , a involved repair, necessary to prevent accumulation lesions marked γ-H2AX. Altogether protection contribute sustained fertility mice, offering promising broad avenues for preservation cancer patients undergoing chemotherapy.

Language: Английский

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Gene therapy with feline anti-Müllerian hormone analogs disrupts folliculogenesis and induces pregnancy loss in female domestic cats

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Feb. 15, 2025

Abstract For female domestic cats, ovariohysterectomy is the only method of inducing permanent infertility. However, hundreds-of-millions free-roaming cats globally highlight necessity for alternative contraceptive approaches. One strategy involves a single injection vector delivering fertility-inhibiting protein lifetime contraception. Recent studies in mice and have identified anti-Müllerian hormone as an excellent candidate this type Here, we leverage our recent characterization molecular mechanisms underlying human synthesis activity, to generate potent feline analogs. Single intramuscular delivery these analogs using adeno-associated viral leads greater than 1000-fold increase levels, which are sustained 9 months. High serum associated with abnormal estrus cyclicity, non-follicular ovarian cyst formation, progressive decline antral follicle numbers, however, few surviving large follicles continue ovulate. Unlike previous studies, supraphysiologic levels do not block conception, although they incompatible maintenance pregnancy. Our findings complexity effects on physiology but confirm that growth factor fertility control cats.

Language: Английский

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Hormonally-regulated KCTD14 Expression Is Higher in Oocytes from Antral Follicles Compared to Those from Preantral Follicles in Mice, with Implications in Fertility.

PubMed, Journal Year: 2025, Volume and Issue: unknown

Published: March 5, 2025

Language: Английский

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Modeling the extension of ovarian function after therapeutic targeting of the primordial follicle reserve

Human Reproduction Update, Journal Year: 2025, Volume and Issue: unknown

Published: May 5, 2025

Abstract BACKGROUND Women are increasingly choosing to delay childbirth, and those with low ovarian reserves indicative of primary insufficiency at risk for sub- infertility also the early onset menopause. Experimental strategies that promise extend duration function in women currently being developed. One strategy is slow rate loss existing primordial follicles (PFs), a second increase, or ‘boost’, number autologous PFs human ovary. In both cases, would be expected lengthened, menopause delayed. This might accompanied by an extended production mature oocytes sufficient quality fertile lifespan. OBJECTIVE AND RATIONALE this work, we consider how slowing physiological aging improve health well-being patients, summarize current state-of-the-art approaches We then use mathematical modeling determine interventions likely influence quantitatively. Finally, efficacy benchmarks should achieved so individuals will benefit, propose criteria could used monitor ongoing different patients as these validated. SEARCH METHODS Current methods estimate size reserve its relationship timing menopausal transition were compiled, publications establishing designed deliver additional identified. OUTCOMES review our understanding consequences reproductive women, compare may POI. provide decay presence therapies PF boost numbers. An interactive online tool provided estimates impact across natural population. Modeling output shows treatments need applied possible many years achieve significant contrast, add occur late relative Combined while boosting numbers (new) offer some benefits delaying WIDER IMPLICATIONS Extending function, perhaps lifespan, on horizon least patients. without such complements helps guide clinical goal. REGISTRATION NUMBER Not applicable.

Language: Английский

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PARP1-catalyzed PARylation of YY1 mediates endoplasmic reticulum stress in granulosa cells to determine primordial follicle activation

Cell Death and Disease, Journal Year: 2023, Volume and Issue: 14(8)

Published: Aug. 15, 2023

Although only a small number of primordial follicles are known to be selectively activated during female reproductive cycles, the mechanisms that trigger this recruitment remain largely uncharacterized. Misregulated activation may lead exhaustion non-renewable pool follicles, resulting in premature ovarian insufficiency. Here, we found poly(ADP-ribose) polymerase 1 (PARP1) enzymatic activity surrounding granulosa cells (GCs) determines subpopulation dormant awakened. Conversely, specifically inhibiting PARP1 oocytes an vitro mouse follicle reconstitution model does not affect activation. Further analysis revealed PARP1-catalyzed transcription factor YY1 PARylation at Y185 residue facilitates occupancy Grp78 promoter, key molecular chaperone endoplasmic reticulum stress (ERS), and promotes GCs, which is required for GCs maintaining proper ERS Inhibiting prevents loss by attenuating excessive under fetal bisphenol A exposure. Together, demonstrate acts as pivotal modulator determine fate represent novel therapeutic target retention females.

Language: Английский

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